Non-Viral Deoxyribonucleoside Kinases – Diversity and Practical Use

Christiansen, Louise Slot; Munch‐Petersen, Birgitte; Knecht, Wolfgang

doi:10.1016/j.jgg.2015.01.003

Cited by 17 publications

(14 citation statements)

References 115 publications

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“…7,9 The ATP-dependent dimer-to-tetramer transition is associated with 30-fold higher catalytic efficiency. 10 The difficulty in describing the structural details of the mechanism of action of hTK1 mainly arises from the difficulty in crystallizing its active state in an open conformation. Thus far, the 3D structure of this enzyme in its active form has not been obtained.…”

Section: ■ Introductionmentioning

confidence: 99%

Inactive-to-Active Transition of Human Thymidine Kinase 1 Revealed by Molecular Dynamics Simulations

Makurat

Cournia

Rak

2021

J. Chem. Inf. Model.

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Despite its importance in the nucleoside (and nucleoside prodrug) metabolism, the structure of the active conformation of human thymidine kinase 1 (hTK1) remains elusive. We perform microsecond molecular dynamics simulations of the inactive enzyme form bound to a bisubstrate inhibitor that was shown experimentally to activate another TK1-like kinase, Thermotoga maritima TK (TmTK). Our results are in excellent agreement with the experimental findings for the TmTK closed-toopen state transition. We show that the inhibitor induces an increase of the enzyme radius of gyration due to the expansion on one of the dimer interfaces; the structural changes observed, including the active site pocket volume increase and the decrease in the monomer−monomer buried surface area and of the number of hydrogen bonds (as compared to the inactive enzyme control simulation), indicate that the catalytically competent (open) conformation of hTK1 can be assumed in the presence of an activating ligand.

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Section: ■ Introductionmentioning

confidence: 99%

Inactive-to-Active Transition of Human Thymidine Kinase 1 Revealed by Molecular Dynamics Simulations

Makurat

Cournia

Rak

2021

J. Chem. Inf. Model.

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“…2.7.1) may answer this need. In fact, dNKs are phosphorylating enzymes which in vivo selectively catalyze the transfer of a phosphate group from a donor nucleotide (typically ATP) to the 5’‐hydroxyl group of a nucleoside thus forming the corresponding 5’‐mononucleotide (Scheme B).…”

Section: Introductionmentioning

confidence: 99%

Immobilization of Deoxyadenosine Kinase fromDictyostelium discoideum(DddAK) and Its Application in the 5’-Phosphorylation of Arabinosyladenine and Arabinosyl-2-fluoroadenine

et al. 2017

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This manuscript is dedicated to the memory of Prof. Jure Piškur Deoxyadenosine kinase from Dictyostelium discoideum (DddAK) phosphorylates its natural substrate (2'-deoxyadenosine, dAdo) as well as the arabinosyladenine analogues vidarabine (araA) and fludarabine (F-araA) to their corresponding 5'-monophosphates. DddAK has been here immobilized by ionic interaction on an aminated epoxy-functionalized support (Sepabeads TM EC-EP), and cross-linked with oxidized dextran. The final activity recovery was 33-42 %, depending on the protein loading. Immobilization enhanced the stability of DddAK at pH 10 and, to a lesser extent, at 45 8C. Phosphorylation of dAdo, araA and F-araA catalyzed by immobilized DddAK was always nearly quantitative in less than 12 hours. Fludarabine monophosphate was synthesized from F-araA (95 % conversion, 20 g/L) using immobilized DddAK in fully aqueous medium, thus showing that this biocatalyst could be used for developing a preparative phosphorylation process greener and more efficient than POCl 3 -based phosphorylation.[a] Dr.

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“…Furthermore, dNKs are able to phosphorylate nucleoside analogs (NA) that are used for the treatment of viral or cancer diseases and this reaction is usually the rate-limiting step [ 1 ]. The diversity of non-viral dNKs and their practical use has been reviewed very recently [ 2 ].…”

Section: Introductionmentioning

confidence: 99%

“…The use of non-viral dNKs in suicide gene therapy has also been reviewed recently [ 2 ]. Among the three non-viral dNKs that have been tested in in vivo animal models so far, we find enzymes from the non-TK1-like group, the ultrafast multisubstrate dNK from Drosophila melanogaster (DmdNK) and the human dCK (HsdCK) [ 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 ].…”

Section: Introductionmentioning

confidence: 99%

New Variants of Tomato Thymidine Kinase 1 Selected for Increased Sensitivity of E. coli KY895 towards Azidothymidine

Christiansen¹,

Egeblad²,

Munch‐Petersen³

et al. 2015

Cancers

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Nucleoside analogues (NA) are prodrugs that are phosphorylated by deoxyribonucleoside kinases (dNKs) as the first step towards a compound toxic to the cell. During the last 20 years, research around dNKs has gone into new organisms other than mammals and viruses. Newly discovered dNKs have been tested as enzymes for suicide gene therapy. The tomato thymidine kinase 1 (ToTK1) is a dNK that has been selected for its in vitro kinetic properties and then successfully been tested in vivo for the treatment of malignant glioma. We present the selection of two improved variants of ToTK1 generated by random protein engineering for suicide gene therapy with the NA azidothymidine (AZT). We describe their selection, recombinant production and a subsequent kinetic and biochemical characterization. Their improved performance in killing of E. coli KY895 is accompanied by an increase in specificity for the NA AZT over the natural substrate thymidine as well as a decrease in inhibition by dTTP, the end product of the nucleoside salvage pathway for thymidine. The understanding of the enzymatic properties improving the variants efficacy is instrumental to further develop dNKs for use in suicide gene therapy.

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Non-Viral Deoxyribonucleoside Kinases – Diversity and Practical Use

Cited by 17 publications

References 115 publications

Inactive-to-Active Transition of Human Thymidine Kinase 1 Revealed by Molecular Dynamics Simulations

Inactive-to-Active Transition of Human Thymidine Kinase 1 Revealed by Molecular Dynamics Simulations

Immobilization of Deoxyadenosine Kinase fromDictyostelium discoideum(DddAK) and Its Application in the 5’-Phosphorylation of Arabinosyladenine and Arabinosyl-2-fluoroadenine

New Variants of Tomato Thymidine Kinase 1 Selected for Increased Sensitivity of E. coli KY895 towards Azidothymidine

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